KR20010089781A - Soy depigmenting and skin care compositions - Google Patents

Abstract

The present invention relates to a skin care composition for topical delivery of soy products, including unmodified soy products and stabilization systems, wherein the stabilization system comprises a component selected from the group comprising antioxidants, chelating agents, or preservatives. Include. The invention also relates to methods of using the skin care compositions of the invention to treat skin diseases and to enhance the appearance and feel of the skin.

Description

Bleaching and skin protection composition using soybeans {SOY DEPIGMENTING AND SKIN CARE COMPOSITIONS}

Understanding the chemical and enzymatic basis of melanogenesis is documented in large quantities. Melanocytes are transferred from the embryonic neural crest to the skin to produce melanin-producing melanosomes, secretory granules. Melanogenesis occurs in melanosomes, which are then distributed to keratinocytes through melanocyte dendrites. The main enzyme in melanogenesis is tyrosinase, which initiates a cascade of reactions that convert tyrosine to biopolymer melanin. Two tyrosinase related proteins (TRPs) are known, TRP-1 and TRP-2. These proteins share about 40% homology with tyrosinase and have a catalytic activity as well as a regulatory role in melanogenesis. TRP-1 is the most abundant glycoprotein in melanocytes.

The coloring of the skin has been a concern for many years. In particular, the ability to remove age spots, freckles, or hyperpigmentation such as found in aging skin is a general concern for individuals who desire uniform complexion. In certain parts of the world, general whitening of the body is desired. Many methods have been proposed to achieve discoloration. For example, kojic acid, hydroquinone, retinoids, and other chemical compounds have been used for bleaching. The majority of the above solutions were found to be unacceptable. Often there is a distinct demarcation between the areas of the skin to which such compositions have been applied. Therefore, accurate application of these compositions is necessary to achieve the required results. Many of these compositions have been found to irritate the skin and their use is undesirable.

Post-inflammatory hyperpigmentation is a frequent problem with various cutaneous disorders along with therapeutic interventions. However, the potential mechanisms and diversity of individuals developing post-inflammatory hyperpigmentation are not well known. Pulsed laser therapy is not effective against intradermal composites or post-inflammatory pigmentation and is not always affordable. Hydroquinones and light-spectrum sunscreens are generally used after treatment, but they do not completely prevent or eliminate excessive pigmented lesions. There is a great need for safe, effective and applicable therapies for post-inflammatory over-pigmentation. What's even more demanding is that these therapies can be incorporated into everyday skin care products to create healthier, more pleasing skin.

Aging of the skin is a complex phenomenon resulting from the interaction of several essential and external factors. Changes in skin associated with aging often appear as cosmetic disorders. Due to psychological effects, aging of the skin has become an issue of immense social significance and concern. With the aging of baby boomers, and in the age of beauty care, beauty maintenance and rejuvenation are becoming increasingly interested. There is a great need for a method of preventing and treating skin aging. Intrinsic aging is inevitable and is a common process. Among the external influences (wind, heat, tobacco smoke, chemicals, etc.), ultraviolet light seems to be the single most important factor associated with skin aging. Photoaging is induced due to cumulative exposure to ultraviolet (UVR). Increasing sun exposure during recreation, including excessive sunbathing, depletion of stratospheric ozone, and the use of UVR in the treatment of various skin diseases have led to more widespread photoaging in the last decade. Photodamage can be prevented by sun protection and adequate sun protection and reversed by the use of topical retinoids, but this is very irritating and expensive. Excessive exposure to ultraviolet and visible light causes sunburn. The use of aspirin and other non-steroidal anti-inflammatory drugs, cool baths and topical steroids only provide pain relief.

There is a need for a single topical treatment that can prevent or reverse skin aging and prevent or alleviate sunburn. As a single and affordable topical treatment for daily skin protection, it can provide these effects without irritation or side effects, additionally provide the desired uniform skin tone, flatten the skin and skin tone, make the skin firm, It is further desirable to have a treatment that leads to a healthy, young look.

Acne is a disease of inflammatory skin that often occurs in adolescence and occurs slightly in older people. Acne status ranges from the comedo of pilosebaceous follicles to more severe como-inflammatory symptoms such as pustules, papules, papules and nodules. May include skin damage. The above conditions are not only uncomfortable for the victim, but can also be embarrassing, detrimental, and terrifying.

Many different approaches to improving these diseases have been in the past, some of which have been more effective than others. Methods ranging from simple face wash and cleansing to medicines have been adopted.

There is a need for a single topical treatment that can prevent or reverse acne and does not require pharmaceutical preparation. There is a further need for a single, affordable topical treatment for daily skin protection, which can provide this effect without irritation or side effects, additionally providing a desirable uniform skin complexion, flattening skin tone and skin tone, It is further desirable to have a treatment that will lead to a healthy and healthy look.

Dimpling of the skin of the thighs and buttocks is a phenomenon commonly referred to as cellulite, which affects women more often than men. The basic pathophysiology of cellulite is not clear. In theory, cellulite may reflect differences in non-involved individuals or portions of individuals and in the adipose tissue biochemistry or connective tissue structures of the affected individuals. Although the connective tissue of the female thigh is configured to highlight differences in small sub-dermal adipose tissue tissue depots, fat tissue physiology, blood flow, or cellulite etiology There is no evidence of any major role in biochemistry. Products aimed at curing cellulite (eg, cellasene) are widely sold around the world, but their efficacy is questionable. Treatment of cellulite, such as metal-probe ultrasonic-assisted lipoplasty techniques or syringe liposculpture, has some consequences on the contouring of the body but is painful and It is expensive and requires repeated treatment. There is a need for a topical and affordable treatment that can prevent or reverse cellulite and make the skin firm, making the skin look healthy and young, and providing daily skin protection without irritation and side effects.

In the applicant's already filed PCT patent application WO 99/04752, a non-pasteurized soybean-derived extract for bleaching the skin is described. Applicants newly discovered a new stable and cosmetically superior composition containing soy products containing up to 0.1% surfactant and less expensive to produce with greater skin depigmentation activity. It was unexpectedly found that soy-containing compositions have anti-aging activity, which increase the elasticity and firmness of the skin, and are useful for the treatment and prevention of sun damage and acne.

The present invention relates to a composition containing a non denatured soybean product that can be used to bleach, smooth skin tone and skin texture, firm skin and protect the skin. Specifically, the present invention is to create a skin having a uniform tone and color, increase the elasticity of the skin, reduce the aging of the skin, make the skin firm and decolorize to promote the youth of the skin, damage to the skin by sunlight To prevent and treat acne by the treatment of cellulite, to control bleeding, and to use soybean-containing compositions to induce aesthetically pleasing skin texture. It is about.

1 is three graphs demonstrating that soy milk can inhibit trypsin, PAR-2 cleavage, and keratinocyte phagocytosis.

FIG. 2 is a histological partial view of pigmented pig skin treated with soy formulation and to demonstrate pigment deposition. FIG.

3 is a photograph of the side of a pig with black skin treated with soymilk and DHLA formulations.

4 is a histological partial view of rat skin treated with soymilk and stained to show elastin fibers.

5 is a photograph of a person's face overexposed to the sun after half the face has been treated with soybean milk.

6 is an enlarged view of the skin of a person overexposed to the sun after half of the face has been treated with soymilk.

7 is a result diagram for showing acne incidence (acnegenicity) and irritancy (irritancy) in the form of a graph.

FIG. 8 shows a Western blot of soybean formulations for investigating soybean Trypsin Inhibitor (STI). FIG.

DETAILED DESCRIPTION The present invention provides skin protection compositions, antioxidants, and chelates for topically delivering soy products, such as soy products (e.g., non-protein denatured soy milk or powder), to mammals such as humans, and the like. A stabilizing system comprising one or more elements selected from the group consisting of first or preservatives. Preferably, the composition of the present invention does not contain more than about 0.1% surfactant. On the other hand, the composition may further include, but is not limited to, one or more other skin-related active agents such as depigmenting agents, thickening agents, emollients, antioxidants or sunscreen agents.

The invention also relates to a method for bleaching the skin of a mammal, the method comprising applying the composition to the skin.

The invention also relates to an improved method for protecting the skin. Application of soymilk or soy-derived compositions to the skin improves the firmness, elasticity, even skin tone and texture of the skin, reduces skin aging, sun-induced damage and acne It has been found to be useful for the prevention and treatment of acne. The present invention provides examples of soy-derivative compositions useful for skin protection.

Other features and advantages of the invention will be apparent from the description and the claims.

Those skilled in the art will be able to fully practice the present invention based on the detailed description of the present invention. The following specific examples are described by way of example only, and are not intended to limit the scope of the invention. Technical and scientific words that are not defined in the specification of the present invention will be commonly understood by those of ordinary skill in the art to which the present invention belongs. In addition, all publications, patent applications, patents, and other references mentioned herein are incorporated by reference in the present invention.

The novel compositions according to the invention comprise legume products, preferably soy products, which may be in the form of a fluid (eg soy milk) or a solid (eg soybean powder or soy milk powder). have. "Bean product" refers to a soy-derived substance that contains ingredients found naturally in soybeans, at the relative concentration conditions found in soybeans. In one embodiment the soy product is an untreated artificial soy product. "Deaturation" means "change of the physical and physiological properties of a protein by heat, X-rays or chemicals," according to the Anti-taming Medical Dictionary (1990 edition). Such changes include decreased activity (if present) and reduced (or altered) the antigenicity of the antigen (in the antigen). A "non-denatured soy product" refers to such soy products (eg in soy milk) to form liquids (such as to form creams, lotions, injectable solutions or gels) based on the composition. Refers to a soy product containing the process of induction. The composition may comprise from about 50 mass% to about 98 mass% (eg, from about 70 mass% to about 98 mass%). The composition according to the invention may be delivered topically, orally, or parenterally, but is not limited to the methods of administration.

Soybean products useful in the present invention may be produced from all soybean species regardless of geographic origin, sun exposure, harvest time and the like. However, certain strains, geographic origins or growth conditions may be more desirable. For example, but not limited to, growth conditions such as soybean strains rich in trypsin inhibitors (eg STI, LTI, BBI) or other legume strains, or trypsin inhibitors in beans may be preferred. Legumes useful in the compositions of the present invention have a distinctive odor, which should be appreciated in certain cultures, but may be undesirable in other cultures. Thus, if necessary, odors of the compositions according to the invention include, but are not limited to, soybeans containing lipoxygenase-2-deficient and soybeans containing a modified sugar profile, and the like. Can be reduced by using soy products derived from certain strains of soy known to produce reduced odors. Reducing the level of oxygen in the formulation can also reduce odor. Various masking agents or fragrances can also be used to mask the odor.

In one embodiment, the composition according to the invention comprises one or more preservatives. Since the composition according to the invention has not been denatured (ie, a composition in which protease inhibitor activity is maintained), it may be further preferred as a medium of microbial growth. Preservatives are useful for substantially preventing the decomposition of microorganisms. Preservatives include, for example, parabens such as methyl-paraben, ethyl-paraben and propyl-paraben and phenoxy ethanol. Other examples of preservatives are described in pages 1654-55 of Weninger and Megwen's International Cosmetic Ingredient Dictionary and Handbook (CTFA, 7th edition, 1997) (hereinafter referred to as "cosmetic handbook"). The composition may comprise from about 0.01% to about 20% by weight (more preferably, from about 0.5% to about 5% by weight). Microbial contamination can also be removed by gamma irradiation or by simple heat treatment that does not eliminate microfiltration or protease inhibitory activity.

Antioxidants and / or chelating agents can be used to increase shelf life and stability of the composition. Antioxidants can be added for both formulation stabilization and biological efficacy. Antioxidant compounds and derivatives thereof include sulfhydryl compounds and derivatives thereof such as sodium metabisulfite and N-acetyl-cysteine, lipoic acid and dihydrolipoic acid, resveratrol, Water soluble antioxidants such as acetyl-cysteine (R) Iniferine or lactoferrin and ascorbic acid and ascorbic acid derivatives such as ascorbyl palmitate and ascorbyl polypeptides. Fat-soluble antioxidants suitable for use in the compositions according to the present invention include butylated hydroxytoluene, retinoids such as retinol and retinyl palmitate, tocopherols such as tocopherol acetate, tocotrienol and ubiquinone It is not limited to this. Natural extracts comprising antioxidants suitable for use in the compositions according to the invention include flavonoids and isoflavonoids and derivatives thereof, such as extracts containing genistein and diadzein, extracts containing resveratrol and the like. Examples of such natural extracts include grape seeds, green tea, pine bark, propolis, and legume extracts, and examples of other antioxidants can be found in the Cosmetic Handbook, pages 1612-13. The composition according to the invention may comprise an antioxidant in an amount from about 0.001% to about 20% by weight (eg, from about 0.01% to about 10% by weight) of the composition.

Chelating agents are also useful in helping to stabilize the compositions of the present invention. Examples of chelating agents include EDTA and its derivatives (eg, disodium EDTA and dipotassium EDTA), iniferine®, lactoferrin, and citric acid. Another example of a chelating agent is listed on page 1626 of the Cosmetic Handbook. The composition of the present invention may comprise a chelating agent in an amount of about 0.001% to about 20% by weight (eg, about 0.01% to about 10% by weight).

Thickening agents (eg thickeners or viscosity increasing agents) can be used to alter the viscosity in the compositions of the present invention. Preferred viscosities of the present compositions depend on the intended use (eg, bath product, cream, lotion, or gel). For example, when used as a bath or cleaning product, the viscosity of the composition should be relatively low, similar to the viscosity of aqueous solutions. When used as a cream, lotion, or gel, the viscosity will be slightly higher (eg, between about 100 cps and about 100,000 cps).

Thickeners that can be added to the compositions of the present invention to change the viscosity include polymers such as polyacrylates (eg, polyacrylamides). Examples of other viscosity-modifying agents are listed on pages 1692-1697 of the Cosmetic Handbook. In order to obtain a suitable viscosity, the compositions of the present invention may comprise from about 0.01% to about 20% by weight (eg, about 0.1% to about 5% by weight) thickener.

The composition of the present invention may comprise other bleaching agents in addition to soy products. Examples of such bleaching agents include lipoic acid, dihydrolipoic acid, resveratrol, ascorbic acid, kojic acid, hydroquinone, and isoflavones. , Retinoids (eg, retinol, retinoic acid and retinyl palmitate), tyrosinase inhibitors, melanosome transfer inhibitors and melanin Selective cytotoxic agents on melanocytes or natural extracts that include these effects include, but are not limited to. The amount of bleach used will vary depending on the activity of the compound and will generally be from about 0.001% to about 20% (eg, from about 0.01% to about 10% by weight) by weight of the composition.

The composition of the present invention may include a compound that enhances the feeling of the composition in contact with the skin of the user. Examples of such compounds include oils, silicones (eg, siloxane polymers such as dimethicone) and skin-conditioning agents such as emollients and humectants. ), But is not limited to such. Examples of such skin conditioning agents can be found on pages 1656-1670 of the Cosmetic Handbook.

Compositions of the present invention may advantageously also include, but are not limited to, other cellulite inhibitors such as caffeine, retinol and other retinoids. Of course, anti-inflammatory drugs such as corticosteroids, antipruritic agents, topical analgesics, acne inhibitors such as benzoyl peroxide and acne such as beta-hydroxy acids such as salicylic acid. Anti-aging products including inhibitors, retinoids, retinol, alpha-hydroxy acid, co-enzyme-Q, miconazole, ketoconazole, erubiol ), Antibiotics and antimycotics and shine-control products, including itraconazole, and other products known to those of ordinary skill in the art. External topical formulations can be mixed into the compositions of the invention and used in the methods of the invention.

The compositions of the present invention can be prepared by mixing the appropriate ingredients. For example, soymilk is mixed with chelating agents, preservatives and / or antioxidants. A thickener is then added to this system and the mixture is further mixed until it is homogeneous to the appropriate viscosity. The composition of the present invention may be prepared under an arcon or nitrogen (or other inert gas) blanket to improve formulation stability and / or reduce the odor of soybeans. Compositions of the present invention include tubes, sealed packets, jars, pumps, bottles, cans, gadgets, towellets, wipes. Or the like. Sealed packages such as aluminum tubes, aluminum pockets, pumps, and laminate tubes can also be used to further improve the stability of the product.

The compositions of the present invention can be used to bleach skin of mammals (eg, humans). Decolorization means lightening the color of the skin area, which is desired because of the user's skin tone / complexion (for example, the entire body, which is not uniform due to aging of the face, hands or skin, or because of race or illness). Darker throughout the body), even skin tone, and darker pigmented areas such as aging spots, freckles, and unrestricted but post-inflammatory hyper-pigmentary lesions. Including but not limited to specifically brightening.

It is unexpectedly found that the compositions of the present invention can be used to treat skin of a person causing reduced skin oiliness and gloss. When applied topically to the skin, including those used as bath additives or in addition to wipes, the compositions of the present invention, including but not limited to, soy-containing compositions comprising protease inhibitory activity, may be used to provide an oily facial skin. Reduces gloss and In one preferred embodiment, the composition may be applied daily at least once a day as long as the user desires the desired effect.

The compositions of the present invention can also be used to prevent or treat acne. The composition of the present invention may be applied preferably twice a day for a period of time, preferably for at least about 45 days, so that it may prove to improve the appearance of the skin and treat the disease. However, depending on the sebaceous gland activity of the individual, it is believed that applying the present composition for at least about 2 to about 4 weeks should provide for amelioration or prevention of the disease.

In another embodiment, soy products containing soybean protease inhibitory activity, but not limited to the compositions of the present invention, when used topically to the skin, including those used as bath additives or contained in patches, soothing, refreshing to the user And / or, it has been found to help relax effects and give an aesthetically pleasing skin feel. The amount and frequency of application of the composition to be applied will depend on the concentration of the soybean agent in the composition (e.g., soy milk), the condition of the skin area being treated, and the desired effect, which ultimately will be determined by the user of the composition. will be. In one embodiment the composition is applied daily at least once a day as long as the user desires a desired effect.

In addition, it was unexpectedly found that applying soymilk to human skin after overexposure to the sun relieves pain and burning sensation caused by sunburn and reduces skin peeling. The present invention provides a method for the calming, refreshing and pain relief effect after sunburn or excessive exposure to the sun. The method consists of soymilk containing a composition containing a protease inhibitory activity, consisting of daily topical application at least once a day as long as the user desires a desired effect.

In another example, it was unexpectedly found that applying soymilk to human skin in advance (eg, used as a bath additive to apply topical application) could prevent sun-induced damage. After applying soymilk, the skin was overexposed to the sun, reddening, peeling, drying, and painless like the unapplied skin of the same person. The present invention provides a method for preventing skin peeling, redness and pain after excessive sun exposure. The method consists of soymilk containing a composition containing a protease inhibitory activity, consisting of daily topical application at least once a day as long as the user desires a desired effect.

Therefore, soymilk and its derivatives have elasticity, prevent skin peeling / redness / pain from overexposure to the sun, refreshing, bathing, discoloring, evening skin tone and tissues, and inflammation after sun exposure / sunburn. Skin protection properties, including post-pigmentation pigmentation, acne and cellulite.

As indicated by the examples herein, it has unexpectedly been found that the topical use of soymilk increases the elasticity of the applied skin. Therefore, proper use of soybean extracts containing protease inhibitory activity and, in particular, soymilk and soybean formulations, may affect elastin and collagen fibers and skin elasticity properties, resulting in increased skin elasticity and firming. do. Topical application of soy products containing protease inhibitory activity, which is not limited to the compositions of the present invention and our previous invention (PCT patent application WO99 / 04752), results in firm skin, increased skin elasticity, and anti-aging effects. Unexpectedly found that aesthetically pleasing skin feel is induced. The present invention provides a method of increasing the firmness and elasticity of the skin and reducing the signs of skin aging, which is useful for whole body parts, preferably the face, upper body and thigh and hip skin. The method consists of soymilk containing a composition containing a protease inhibitory activity, consisting of topical application at least once daily, as long as the user desires a desired effect.

The following examples illustrate some examples of the compositions of the present invention. They should not be used to limit or limit the scope of the invention in any way.

Example 1 Preparation of Soy Milk from Soybean Powder

Add 160 grams of soybean powder (available from Sunlight Foods, Taipei, Taiwan) to about 1440 grams of deionized water. The mixture is stirred at room temperature for about 1 hour. The mixture is then filtered through a sieve having a pore diameter of 75 μm. Filtration yields about 1.1 kg of soymilk.

Example 2 Preparation of Soymilk Gel from Soymilk

The composition described below of the present invention is prepared as follows. The weight percent of each component in the composition is shown in Table 1 and Table 2 below. First, the soymilk prepared in Example 1 is placed in the first beaker. Preservatives or preservatives phenoxyethanol sold under the trademark Phenonip (a mixture of preservatives methyl-paraben, propylene-paraben, ethyl-paraben and phenoxy-ethanol, available from NIPA, Wilmington, DE) Add to soy milk. Next, the chelating agent Disodium EDTA and the humectant glycerin in various examples are placed in a first beaker and mixed with soymilk. Cyclomethicone, or dimethicone (trade name Dow Corning 200 Fluid), or Polysorbate 20, or Aluminum Starch Octyl Succinate, or Sucrose Cocoate ), Or additional addition of PEG-6 Capric / Caprylic Triglycerides to the soymilk mixture at this stage as required in the various examples in Tables 1 and 2 It is possible. The concentrate of claim polyacrylamides (polyacrylamide), Laureth -7 (laureth-7) and C _13-14 iso-paraffins (C _13-14 isoparaffin) (available obtained from Seppic Paris, France registered trademark Sepigel) antioxidants BHT and Put together in a second beaker. Then add the ingredients in the second beaker to the ingredients in the first beaker and mix until uniform. The antioxidant ascorbic acid, sodium ascorbyl phosphate, lactoferrin, or tocopherol is then added to the beaker and mixed uniformly to produce the final gel.

Example 3 Preparation of Soymilk Gel from Soybean Powder, Soybean Milk Powder or Soybean Extract

The composition described below of the present invention is prepared as follows. The weight percentages of each component in the composition are shown in Table 2 below. First, soybean powder (available from Devansoy Farms, LA Carroll) or soybean powder (available from Sunlight Foods, Taipei) or soybean extract; Place deionized water in the first beaker and mix to reconstitute soybean powder. The preservative and chelating disodium EDTA sold under the trademark Phenonip are then added to the first beaker and mixed with soymilk. The polyacrylamide, laureth-7 and C _13-14 isoparaffin mixture is placed in a second beaker with the antioxidant BHT. Then add the ingredients in the second beaker to the ingredients in the first beaker and mix until uniform.

Soybean Essence One 6 8 21 23 Soymilk 87.42% 89.04% 96.09% 96.05% 95.70% Phenoxyethanol 0.73% Phenoxyethanol and Parabens 1.00% 1.00% 1.00% 1.00% Glycerin 2.50% 2.50% Cyclomethicone 2.00% Aluminum Starch Octyl Succinate 0.75% Sucrose Cocoate 1.00% 1.00% PEG-6 Capric / Caprylic Triglycerides 3.00% 3.00% Disodium EDTA 0.10% 0.10% 0.05% Polyacrylamide / Laureth-7 / C _13-14 Isoparaffin 2.50% 2.75% 2.90% 2.90% 3.20% Ascorbic acid 0.01% Butylated Hydroxytoluene 0.10% 0.01% 0.05% 0.05% Polysorbate 20 0.50% sum 100% 100% 100% 100% 100%

24 26 27 28 33 34 35 Soy milk 94.40% 92.40% 90.70% 94.70% Phenoxyethanol and parabens 1.00% 1.00% 1.00% 1.00% 1.00% 1.00% 1.00% glycerin 5.00% Disodium EDTA 0.05% 0.05% 0.05% 0.05% 0.05% 0.05% 0.05% Polyacrylamide / Laureth-7 / C _13-14 Isoparaffin 3.50% 3.50% 3.20% 3.20% 3.20% 3.20% 3.20% Ascorbic acid 1.00% Butylated Hydroxytoluene 0.05% 0.05% 0.05% 0.05% 0.05% 0.05% 0.05% Deionized Water 90.70% 90.70% 85.70% Lactoferrin 1.00% 1.00% Tocopherol 1.00% Dow Corning 200 Fluid 1.00% Soy milk powder 5.00% Soybean Extract with Ethanol / Water Mixture 5% 10% sum 100% 100% 100% 100% 100% 100% 100%

Example 4 Soymilk Interfering with the PAR-2 Pathway

Applicant's patent application WO 99/04752 describes the importance of the PAR-2 pathway in staining and also states that the protease inhibitor STI induced by soybeans interferes with this pathway and causes discoloration. have. In addition, Applicant's patent application WO 99/30729 describes the role of the PAR-2 pathway in inducing keratinocyte phagocytosis and the effect of the PAR-2 pathway on melanosome transfer and staining. In order to test the possibility that soymilk may cause discoloration by inhibiting PAR-2 induced phagocytosis, the following experiments were conducted. First, STI and soymilk act as serine protease inhibitors by inhibiting trypsin-induced cleavage of fluorescent casein peptides in a dose-dependent manner (FIG. 1A). Seems to do. Total protease activity is measured using the EnzChek® Protease Assay Kit, which is the manufacturer's instructions (molecular probes) described below. STI and soymilk are diluted in PBS (Life Technologies) and incubated at 1: 1 (v / v) with 1000 units of trypsin prepared in PBS. After incubation for 1 hour at room temperature using a BODIPY fluorescent casein incubator, the fluorescence was measured on a trademark SpectraMax Gemini microtiter plate reader (Molecular Devices Corporation) using the trademark SoftMax Pro 3.0 software (Molecular Devices Corporation) (here 505 / Radiation 515). Each experiment was performed on six replicates and repeated three times. The percent inhibition of trypsin cleavage of the substrate by STI and soymilk was calculated using Microsoft Excel and graphed by Sigmaplot. As can be seen in FIG. 1A, soymilk can inhibit cleavage due to trypsin in a dose response manner.

In the second experiment, a similar assay was designed to test the specific inhibition of soymilk against PAR-2 cleavage. In this essay, SKGRSLIGK, a human PAR-2 (similar essays are described in the following books: Lourbakos A, Chinni C, Thompson P, Potempa J, Travis J, Mackie EJ, Pike RN Cleavage and Activity of Proteinase-Activated Receptor-2 on Human Neutrophils by Ginger Pain-R Synthesis Including Cleavage Sites of FEBS Lett 435: 1, 45-8, Sept. 11, 1998 Fluorescent peptides replace trypsin with casein peptides as substrates. Peptides are labeled with fluorophore paireds / dabsil (available from Advanced Bioconcept of Montreal, Canada) and are used as substrates for PAR-2 serine protease activators. Cleavage of the peptide with a PAR-2 activator such as trypsin can be detected as fluorescent (excitation 335 / radiation 515). Inhibition of PAR-2 peptide cleavage caused by trypsin by STI or soymilk has trypsin [at 100 mM TRIS (hydroxymethyl) aminomethane buffer, pH 8.0, Digene, Beltsville, MD] 10 units, with or without test substance And 100 micromolar peptides are measured by incubating for 1 hour at room temperature blocked from light. Fluorescence was measured on a trademark SpectraMax Gemini microtiter plate reader (Molecular Devices Corporation) using the trademark SoftMax Pro 3.0 software (Molecular Devices Corporation). Each experiment was performed on six replicates and repeated three times. The percent inhibition of trypsidism was calculated using Microsoft Excel and graphed with a Sigma plot. As can be seen in FIG. 1B, cleavage of the PAR-2 peptide by trypsin was completely inhibited in the presence of STI. Soymilk with the ability to inhibit PAR-2 cleavage resulting from the trypsin (FIG. 2B) suggests that soy extract can inhibit PAR-2 activity similar to STI.

To investigate the possibility that soymilk can inhibit keratinocyte phagocytosis induced by PAR-2, we used the Vibrandt Phagocytosis Assay (Molecular Probes). HaCaT keratinocytes were treated with SLIGRL, PAR-2 for activating peptides and increasing the concentration of STI or soymilk for 24 hours, followed by 100 μl of fluorescently-labeled E.coli K-12 bioparticles. Incubated for hours. Intracellular fluorescence showing the ingested E. coli particles was measured after trypan blue quenching of unabsorbed fluorescence with trypan blue. Fluorescence was measured (485 nm excitation / 538 nm emission) using a SpectraMax® Gemini microtiter plate reader (Molecular Devices Corporation, Sunnyvale, Calif.). Data was collected using Softmax® Pro 3.0 software (Molecular Device Corporation) and SigmaPlot® 5.0 (SPSS Science, Chicago, IL) and SigmaStat® This was analyzed using 2.0 (SPSS Science) software. Each experiment was performed on six replicates, which was repeated at least three times. As shown in FIG. 1C, SLIGRL increased keratinocyte phagocytosis and STI inhibited this increase in a dose dependent manner. Interestingly, freshly prepared soymilk was able to similarly inhibit SLIGRL, which causes phagocytosis in keratinocytes. These three experiments show that soymilk contains protease inhibitory activity, which inhibits PAR-2 activation and eventually reduces keratinocyte phagocytosis. This data indicates that the depigmentation activity of soy products is partly due to their PAR-2 inhibitory activity.

Example 5 Depigmentation Activity of New Soymilk Formulations-Excellent

Our patent application WO 99/04752 shows a soymilk-containing formulation for use in depigmentation and depigmentation of soymilk. One example of such a formulation is Soy Essence 1, described in Table 1 above. The present invention discloses a depigmentation composition comprising a soy product and a stabilizing system containing only 0.1% of a surfactant. One example of such a formulation is the soy essence 23 described in Table 1 above. The following experiments were conducted to demonstrate that soy essence 23 is superior in depigmentation activity than essence 1.

Yucatan microswine (Charles River, Maine) of black skin was placed in an appropriately sized cage in an environmentally controlled location with a light and dark photoperiod of 12 hours each, to provide food and water randomly. Animal care was based on "Guidelines for the Care and Use of Laboratory Animals" (NIH Publication 85-23). 20 μl of test compound was applied topically to the dorsal part of the pig twice daily, 5 days / week, for 8 or 9 weeks. For each pig, one side was always treated from head to tail and the other from tail to head. Biopsies were performed according to standard methods. All pig studies showed no visible inflammation, and biopsies showed no signs of inflammation or other pathological signs.

A portion of the biopsy tissue was stained with Fontana-Mason (F & M) according to standard experimental procedures (Sheenan and Hrapckak, 1980). F & M staining identifies and displays silver nitrate which reduces the molecule. This nonspecific staining in the skin mainly represents melanin. At least three sections were treated per living tissue. Each experiment was repeated at least three times.

2 shows tissue sections from the two experiments. The sections were stained with F & M to indicate melanin deposition in the treated areas. The top shows portions of a pig treated with control (a) and soy essence 23 (b). Bottom shows part of another pig treated with control (c) and soy essence 1 (d). From this figure it is clear that both the relative and absolute reduction in pigmentation induced by soy essence 23 is superior to the level induced by soy essence 1.

Example 6 Depigmentation Activity of Soymilk Promoted in Combination with Antioxidants

Our patent application WO 99/04752 shows the depigmentation activity of soymilk. As can be seen, this depigmentation activity is due in part to the inhibition of keratinocyte phagocytosis induced by PAR-2. To test the hypothesis that improved depigmentation is made by combining one or more mechanisms leading to depigmentation, we tested the depigmentation effect of soymilk and antioxidant combinations.

Pigs are treated twice daily with soy milk or soy milk essence formulations, 1% dihydrolipoic acid (DHLA), and 1% Resveratrol (Res), or soy milk once a day, and DHLA Or Res was treated once a day in combination. A noticeable whitening of the skin was observed after 7-8 weeks of treatment. One example of such an experiment is shown in FIG. 3 showing black skin pig flank flesh treated with a combination of soymilk essence and DHLA and eight weeks old. The left part 1 was treated with soymilk essence 23, the soymilk formulation described above. The right part (3) was treated with DHLA and the middle part (2) was treated with both soymilk essence 23 and DHLA. This figure clearly shows that the combination of soymilk essence 23 and DHLA enhanced depigmentation compared to the treatment of each element alone. Similar results were obtained when using soymilk or soymilk essence in combination with DHLA or Res. This example clearly shows that additives of different mechanisms can enhance the depigmentation effect of soymilk.

Example 7 Soy Milk Affects Skin Elasticity

During the soymilk study described in our applications JBP-430, JBP-464 and JBP-467, we unexpectedly touched and found that the skin of rats treated with soymilk was more elastic. To test the possibility that soymilk has a positive effect on skin elasticity, we conducted two sets of experiments: non-invasive elasticity measurement (cutometer measurement) and staining of skin sections on elastin fibers. In these studies, soymilk treatment increases the elastin fibers of the skin and increases the elasticity of the treated skin.

C3H and C57B1 / 6 mice were induced with a new hair cycle or hair removal as described in our applications JBP-430, JBP-464 and JBP-467. Mice were treated with soybean milk daily as described in the application. A cutometer measurement was performed to measure skin elasticity on the first day of the experiment (day 1, before the first treatment), and on day 21. We used a cutometer available from Acaderm (Menlo Park, California), and in Couturaud et al., "Skin Biomechanical Properties: In Vivo Evaluation of Influence of Age and Body Site by a Non-Invasive Method" {1 Skin Res. and Technol. 68-73 (1995)} and Elsner et al., “Mechanical Properties of Human Forearm and Vulvar Skin” {122 Br. J. Dermatol. 607-614 (1990). Both of these documents are incorporated herein by reference in their entirety. Suction was performed through a 2 mm hole and the recovery rate after removal of the corresponding displacement and negative pressure was measured. In studies in humans, the ratio of deformation parameters Ua / Uf {skin fatigue rate, ie total recovery from load}, Ur / Uf (biological modulus, ie elastic recovery after load), and Ur / Ue The improvement in (advance rate, i.e., improvement in the deformation resistance of the skin) indicates that the tonicity and elasticity of the skin are better. The deformation parameters Ue, Uf, Ua, and Ur depend in part on the skin thickness. In conclusion, Barel et al., "Suction Method for Measurement of Skin Mechanical Properties: The Cutometer," and a guide to non-invasive methods, and the entirety of Skin 335-340 (1995), are incorporated by reference. Likewise, ratios were used for evaluation.

One example of such an experiment is shown in Table 3 and Table 3 shows that these parameters are improved (which reflects improved skin elasticity) during soymilk treatment. While the variation between animals was significant, the increase in cutometric properties was constant, indicating that soymilk has a beneficial effect on skin elasticity.

Mechanical Properties of Haired, Soymilk-treated C3H Rat Skin Biological physical parameters Day 1 Day 21 Untreated Control Soymilk Processing Untreated Control Soymilk Processing Ua / Uf 0.860 +/- 0.066 0.815 +/- 0.154 0.839 +/- 0.068 0.904 +/- 0.043 Ur / Ue 0.943 +/- 0.172 0.964 +/- 0.103 0.617 +/- 0.151 1.402 +/- 0.846 Ur / Uf 0.720 +/- 0.115 0.677 +/- 0.168 0.506 +/- 0.136 0.787 +/- 0.067

To further study this elasticity effect, skin biopsies were conducted throughout this experiment, and skin sections were fully incorporated by reference, Kligman, LH's "Luna's Technology, Wonderful coloring for elastin "[3 (2) The Amer. J. of Dermatopathol. 199-200 (1981), for elastin according to the method disclosed.

As shown in FIG. 4, elastin fibers around the skin appendage and upper dermis of C3H rats (right panel) treated with soymilk as compared to the untreated control (left panel). The thickness and density of (tinted in dark blue) increased. This increase was detected on day 4 of starting treatment and was constant throughout the study. The same result was also obtained using C57B1 / 6 mice. This example demonstrates that using soymilk and soymilk formulations for skin protection increases the elasticity of the skin and reduces skin aging.

In individual experiments, the effect of soybean powder on collagen synthesis was studied in vitro. The effect of soybean powder was evaluated by assessing the binding of radioactive proline in extracellular collagen three days after treatment to increase collagen synthesis rate in normal human dermal fibroblasts. As shown in Table 4, soybean powder improved the synthesis rate of collagen by 58% in normal human dermal fibroblasts at 1 μg / ml. At lower concentrations, soy powder was not used to show any significant irritation. Thus, the skin is firmer and fuller, and looks younger.

Effect of Soybean Powder on Extracellular Collagen Synthesis in Normal Human Dermal Fibroblasts. Soja (µg / ml) Control 0.01 0.1 One Collagen Synthesis (dpm / cell) * 1000 11.26 ± 1.62 15.0 ± 1.46NS 13.68 ± 2.51NS 17.80 ± 2.01p <0.05 Stimulation + 33% + 21% + 58%

NS: Not noticeable

Example 8: Soymilk Prevents Sun-induced Damage

One person was treated with the composition according to the invention once a day, immediately after shaving, containing soy milk in the right half of the face. Eight weeks after soymilk treatment, the man was exposed to excessive sun light during his leisure activities without the use of sunscreen. This person's skin was "burned". This person's face turned red and painful, and skin scaliness began two and three days after sun exposure. Unexpectedly, the right side of the face pretreated with the soymilk composition according to the present invention did not turn red, and was not painful or flaky. 5 shows two sides of a person's face. In this figure the difference between the redness of the skin and the damage caused by the sun is evident. FIG. 6 shows a high-magnification hi-scope image of human skin, demonstrating skin peeling of the untreated side and smoothing of the treated side. This embodiment provides that the routine use of the soymilk composition according to the present invention can protect the skin from unexpected strong sun exposure and reduce the redness, peeling and pain associated with “sunburn”. Prove that there is.

Example 9: Soymilk That Can Treat Damage Caused by Sunlight

The person described in Example 7 also used a soymilk composition in portions of cotton that were not treated only 24 and 48 hours after exposure to sunlight. Applying soymilk after exposure to the sun's rays does not only moisturize damaged skin. It has been found to reduce the redness and peeling of skin while causing coolness and calming of emotions, alleviating the pain associated with excessive sun exposure, and leaving a "fresh" feeling. These examples demonstrate that topical application of soymilk or soymilk formulations can alleviate pain and reduce redness and skin peeling resulting from excessive sun exposure. If you use soy milk on sun-exposed skin, it will be cool and refreshing.

Example 10: Compositions Containing Soybeans That Can Reduce Acne Lesions

To study the effects of applying soy-containing compositions twice daily for 45 days to subjects with mild acne (ie, having more than 10 non-inflammatory and inflammatory lesions at the start of the study) and those without acne For this purpose, a clinical study was conducted. Calculation of the number of acne lesions, evaluation of erythema and photographic evaluation were performed. Soy essence 23 containing 0.005% isoflavones was applied to the subjects in both groups.

In mild acne subjects, there was a very large decrease in the number of inflammatory papules (p = 0.001) from the baseline, ie 41.9%. There was also a trend towards direct reduction of erythema (p = 0.063) from baseline. In subjects without acne, there was no significant increase in comeedones, papules or pustules. This reduced number of papules is shown graphically in FIG. 7.

Example 11 Soymilk Containing Composition of the Invention Containing Intact STI

The composition containing soymilk of the present invention is non-denatured and has protease inhibitory activity. Such compositions are specific because they contain intact porteains, specifically intact STI, serine protease inhibitors. To demonstrate this specificity, the same volume of essence 23 described in Example 3 above and a commercially available soybean-based product (herenna Rubinstein claiming to have "soja protein" in its ingredients) Future White Essence], for STI content, was compared using a western blot according to Bonifacino JS (ed.) (1999). Immunofluoresence staining. In the current protocol of cell biology on CD-ROM [Teton Data Systems, Jackson, WY], the samples were prepared in RIPA buffer [1% nonidet-P40] dissolved in PBS. , 0.5% sodium deoxycholate, 0.1% sodium docedyl sulfate, and a Complete Protease Inhibitor (Berlinger Mannheim, Indianapolis, Indiana). RIPA lysate (10 μg per lane) was electrophoresed on a 10% SDS-PAGE gel and the protein was enhanced chemiluminescence (ECL) specific protein detection assay (Illinois, Amersham, Arlington Heights). Antibiotics for STI were purchased from Chemicon and used at a 1: 500 dilution.

As shown in FIG. 8, the soy formulation essence 23 contains an intact STI that is the same size as the STI label (moving in parallel lanes). In contrast, both the placebo of Essence 23 and products based on commercially available soy do not contain intact STIs.

While the present invention has been described in conjunction with the description, it is to be understood that the above description is intended to explain the scope of the invention as defined by the appended claims and is not intended to be limiting. Other aspects, advantages and modifications are within the claims.

Claims (13)

As a skin care composition for topical delivery of soy products including unmodified soy products and stabilizing systems, The stabilization system comprises a member selected from the group comprising antioxidants, chelating agents, or preservatives. The skin care composition of claim 1, wherein the composition does not comprise greater than about 0.1% of a surfactant. The skin protection composition of claim 1, wherein the composition comprises an antioxidant. The skin protection composition of claim 1, wherein the composition comprises a chelating agent. The skin protection composition of claim 1, wherein the composition comprises a preservative. The skin protection composition of claim 3, wherein the antioxidant is a component selected from the group comprising BHT. The skin protection composition of claim 4, wherein the chelating agent is selected from the group comprising EDTA and the EDTA derivative. The skin protection composition of claim 5, wherein the preservative is paraben. The skin protection composition of claim 1, wherein the stabilization system comprises BHT, EDTA, or derivatives thereof, and parabens. The skin care composition of claim 1, wherein the unmodified soy product is soymilk. The skin protection composition of claim 1, wherein the unmodified bean product is a soy bean powder. The skin care composition of claim 10, wherein the composition further comprises a thickening agent. A skin care composition for topical delivery of soy products comprising essentially unmodified soy products and stabilization systems, The stabilization system comprises one or more components selected from the group comprising antioxidants, chelating agents, or preservatives.